Hepatitis C virus (HCV) establishes a chronic infection in a high percentage of infected individuals and is associated with progressive liver pathology, including cirrhosis and hepatocellular carcinoma. Antiviral drugs such as interferon alpha and ribavirin have had limited success in controlling HCV infection. As a result, it has become the leading cause for liver transplantation in the US. The HCV polyprotein comprises, from the amino terminus to the carboxy terminus, the core protein (C), the envelope proteins (E1 and E2), p7, a membrane bound protein, whose function is unknown and the non-structural proteins (NS2, NS3, NS4A, NS4B, NS5A and NS5B) which are believed to be important for replication. C codes for the core nucleocapsid protein, E1 and E2 are envelope proteins that coat the virus, NS2, NS3 and NS4A are involved in proteolytic processing of the HCV polyprotein, and NS5B has RNA polymerase activity. The functions of NS4B and NS5A are unknown.
Hepatitis C virus is a significant cause of morbidity and mortality, infecting over 100,000,000 people worldwide. Annual HCV related costs in the United States are about $1 billion. Current therapies are clearly inadequate; the best available treatment at present is the combination of interferon and ribavirin, a treatment which is inconveniently lengthy as it typically lasts over one and a half years, difficult to tolerate in that most patients have flu-like symptoms, and extremely expensive as the cost is in the range of thousands of dollars annually. Not only does the present treatment have these disadvantages, but it is also not particularly effective.
Certain interactions of viral proteins with cell membranes have previously been described. For example, in poliovirus and Hepatitis A virus, the nonstructural protein 2C contains a membrane associating amphipathic helix (See Teterina, N. L., et al., J. Virol. (1997) 71:8962-8972 (poliovirus); and Kusov, Y. Y., et al., Arch. Virol. (1998) 143:931-944 (Hepatitis A). This membrane association appears to play a role in RNA synthesis in poliovirus (Paul, A. V., et al., Virol. (1994) 199:188-199). Replication complexes are localized on the host endoplasmic reticulum (ER) and Golgi in the case of poliovirus (Bienz, K., et al., J. Virol. (1992) 66:2740-2747), and infection with poliovirus induces rearrangements of membranes derived from host ER and Golgi (Schlegel, A., et al., J. Virol. (1996) 70:6576-6588).
It is also known that the Hepatitis C nonstructural 5A protein illustrated below is a potent transcriptional activator (Kato, N., et al., J. Virol. (1997) 71:8856-8859); that amino terminal deletion mutants of Hepatitis C virus nonstructural protein NS5A function as transcriptional activators in yeast (Tanimoto, A., et al., Biochem. Biophys. Res. Commun. (1997) 236:360-364); and that this nonstructural protein physically associates with p53 and regulates p21/Waf1 gene expression in a p53 dependent manner (Majumder, M., et al., J. Virol. (2001) 75:1401-1407).
It has also been reported that a number of positive strand RNA viruses, like HCV, replicate in association with cytoplasmic membranes, although the precise manner of such association and replication is not understood. See, for example, Lazarus, L. H., et al., J. Gen. Virol. (1974) 23:213-218 (foot and mouth disease); Bienz, K., et al., Virol. (1980) 100:390-399 (polio); Froshauer, S., et al., J. Cell. Biol. (1988) 107:2075-2086 (alphavirus); Chu, P. W., et al., Arch. Virol. (1992) 125:177-191 (Kunjin virus); Rice, C. M., in Fields Virology, Fields, B. N., et al., Ed. (1996) Lippincott-Raven Publications: Philadelphia, Pa., pages 931-959 (Flaviviridae).
It is also known that NS5A, the nonstructural Hepatitis C protein used for illustration below is associated with cell membranes (Selby, M. J., et al., J. Gen. Virol. (1993) 74:1103-1113; Hijikata, M., et al., Proc. Natl. Acad. Sci. USA (1993) 90:10773-10777; Moradpour, D., et al., Hepatol. (1998) 28:192-201). This protein, NS5A, also has been reported to interact with other host cell proteins such as PKR protein kinase (Gale, M. J. J., et al., Virol. (1997) 290:217-227) and a SNARE-like protein (Tu, H., et al., Virol. (1999) 263:30-41). NS5A has also been implicated in determining the response to interferon therapy in some groups of patients (Gale, M. J. J., et al., Virol., supra); Enomoto, H., et al., N. Engl. J. Med. (1996) 334:77-81.
There exists a need in the art for compositions, including peptide therapeutics, and methods employing the same, to prevent or inhibit infections due to Hepatitis C Virus, including inhibiting replication and/or pathogenesis due to HCV, with minimal or no adverse side effects.